3D printed scaffolds of biosilica and spongin from marine sponges: analysis of genotoxicity and cytotoxicity for bone tissue repair.

Biosilica (BS) and spongin (SPG) from marine sponges are highlighted for their potential to promote bone regeneration. Moreover, 3D printing is introduced as a technology for producing bone grafts with optimized porous structures, allowing for better cell attachment, proliferation, and differentiation. Thus, this study aimed to characterize the BS and BS/SPG 3D printed scaffolds and to evaluate the biological effects in vitro. The scaffolds were printed using an ink containing 4 wt.% of sodium alginate. The physicochemical characteristics of BS and BS/SPG 3D printed scaffolds were analyzed by SEM, EDS, FTIR, porosity, evaluation of mass loss, and pH measurement. For in vitro analysis, the cellular viability of the MC3T3-E1 cell lineage was assessed using the AlamarBlue® assay and confocal microscopy, while genotoxicity and mineralization potential were evaluated through the micronucleus assay and Alizarin Red S, respectively. SEM analysis revealed spicules in BS, the fibrillar structure of SPG, and material degradation over the immersion period. FTIR indicated peaks corresponding to silicon oxide in BS samples and carbon oxide and amine in SPG samples. BS-SPG scaffolds exhibited higher porosity, while BS scaffolds displayed greater mass loss. pH measurements indicated a significant decrease induced by BS, which was mitigated by SPG over the experimental periods. In vitro studies demonstrated the biocompatibility and non-cytotoxicity of scaffold extracts. .Also, the scaffolds promoted cellular differentiation. The micronucleus test further confirmed the absence of genotoxicity. These findings suggest that 3D printed BS and BS/SPG scaffolds may possess desirable morphological and physicochemical properties, indicating in vitro biocompatibility.

Association of a Skin Dressing Made With the Organic Part of Marine Sponges and Photobiomodulation on the Wound Healing in an Animal Model.

The present study aims to characterize and to evaluate the biological effects of a skin dressing manufactured with the organic part of the Chondrilla caribensis marine sponge (called spongin-like collagen (SC)) associated or not to photobiomodulation (PBM) on the skin wound healing of rats. Skin dressings were manufactured with SC and it was characterized using scanning electron microscopy (SEM) and a tensile assay. In order to evaluate its biological effects, an experimental model of cutaneous wounds was surgically performed. Eighteen rats were randomly distributed into three experimental groups: control group (CG): animals with skin wounds but without any treatment; marine collagen dressing group (DG): animals with skin wounds treated with marine collagen dressing; and the marine collagen dressing + PBM group (DPG): animals with skin wounds treated with marine collagen dressing and PBM. Histopathological, histomorphometric, and immunohistochemical evaluations (qualitative and semiquantitative) of COX2, TGFß, FGF, and VEGF were done. SEM demonstrates that the marine collagen dressing presented pores and interconnected fibers and adequate mechanical strength. Furthermore, in the microscopic analysis, an incomplete reepithelialization and the presence of granulation tissue with inflammatory infiltrate were observed in all experimental groups. In addition, foreign body was identified in the DG and DPG. COX2, TGFß, FGF, and VEGF immunostaining was observed predominantly in the wound area of all experimental groups, with a statistically significant difference for FGF immunostaining score of DPG in relation to CG. The marine collagen dressing presented adequate physical characteristics and its association with PBM presented favorable biological effects to the skin repair process.

Flounder fish (Paralichthys sp.) collagen a new tissue regeneration: genotoxicity, cytotoxicity and physical-chemistry characterization.

Collagen dressings have been widely used as effective treatments for chronic wounds acting as barrier, protecting the area from infections and participating in the healing process. Collagen from fish skin is biocompatible, presents low immunogenicity and is able of stimulating wound healing. In this scenario, skin of flounder fish (Paralichthys sp.) may constitute a promising source for collagen. Then, our hypothesis is that fish collagen is able of increasing cell proliferation, with no cytotoxicity. In this context, the aim of the present study was to investigate the physicochemical and morphological properties of collagen using scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDS), mass loss and pH. Moreover, the cytotoxicity and genotoxicity of collagen were studied using in vitro studies (cell viability, comet assay and micronucleus assay). Fish collagen showed no variation of pH and mass weight, with characteristic peaks of collagen in FTIR. Furthermore, all the extracts presented cell viability at least over 50% and no cytotoxicity was observed. Regarding genotoxicity data, the results showed that only the extract of 100% showed higher values in comparison with negative control group for CHO-K1 cell line as depicted by comet and micronucleus assays. Based on the results, it is suggested that fish collagen is biocompatible and present non-cytotoxicity in the in vitro studies, being considered a suitable material for tissue engineering proposals.

3D Printed Scaffolds Manufactured with Biosilica from Marine Sponges for Bone Healing in a Cranial Defect in Rats.

The inorganic part of marine sponges, called Biosilica (BS), presents an osteogenic potential and the ability of consolidating fractures. Moreover, 3D printing technique is highly effective for manufacturing scaffolds for tissue engineering proposals. Thus, the aims of this study were to characterize the 3D rinted scaffolds, to evaluate the biological effects in vitro and to investigate the in vivo response using an experimental model of cranial defects in rats. The physicochemical characteristics of 3D printed BS scaffolds were analyzed by FTIR, EDS, calcium assay, evaluation of mass loss and pH measurement. For in vitro analysis, the MC3T3-E1 and L929 cells viability was evaluated. For the in vivo evaluation, histopathology, morphometrical and immunohistochemistry analyses were performed in a cranial defect in rats. After the incubation, the 3D printed BS scaffolds presented lower values in pH and mass loss over time. Furthermore, the calcium assay showed an increased Ca uptake. The FTIR analysis indicated the characteristic peaks for materials with silica and the EDS analysis demonstrated the main presence of silica. Moreover, 3D printed BS demonstrated an increase in MC3T3-E1 and L929 cell viability in all periods analyzed. In addition, the histological analysis demonstrated no inflammation in days 15 and 45 post-surgery, and regions of newly formed bone were also observed. The immunohistochemistry analysis demonstrated increased Runx-2 and OPG immunostaining. Those findings support that 3D printed BS scaffolds may improve the process of bone repair in a critical bone defect as a result of stimulation of the newly formed bone.

Fish collagen for skin wound healing: a systematic review in experimental animal studies.

Collagen extracted from fishes has been appearing as an alternative for commercial porcine and bovine collagen and it has been considered interesting especially for membrane manufacturing in tissue engineering. Despite the positive in vitro effects of fish collagen membranes, there is still no understanding of all the benefits that this natural biomaterial plays in the wound healing process, due to the lack of compilation of the results obtained in animal studies. In this sense, the purpose of this study was to perform a systematic review of the literature to examine the effects of fish collagen membranes for skin wound healing in experimental models of skin wound. The search was carried out according to the orientations of Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA), and the descriptors of the Medical Subject Headings (MeSH) were defined: "fish," collagen," "skin," and "in vivo". A total of 10 articles were retrieved from the databases PubMed and Scopus. After the elegibility analyses, this review covers the different origins of fish collagen reported in the different papers from the beginning of 2015 through the middle of 2021. The results were based mainly on histological analysis and macroscopic evaluation, and fish skin collagen was responsible for improving the wound healing rate and the process of reepithelization and collagen deposition. In conclusion, fish skin collagen has shown positive results in in vivo studies and may be a potential biomaterial in tissue engineering.